• Journal of Microbiology and Biotechnology
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• 제5권1호
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• pp.26-30
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• 1995

The cytoplasmic endo-$\beta$-l, 4-glucanase (endoglucanase) was purified from cell extracts of Escherichia coli (pBS1) transformant carrying the Bacillus subtilis endo-$\beta$-l, 4-glucanase gene after full growth, and its molecular weight was found to be 52 kilodaltons (kDa). The endo-$\beta$-l, 4-glucanase isolated from the periplasmic space was smaller than 52-kDa cytoplasmic enzyme. The 52-kDa endoglucanase was found to be cleaved in the periplasm and finally converted to 34.5-kDa protein. Small amounts of both 52-kDa and 34.5-kDa proteins were secreted into the culture broth. The cleavage took place in the C-terminal portion of the enzyme. The N-terminal amino acid residues of both 52-kDa and 34.5-kDa enzymes were determined to be the same, Ala, the 30th residue of the primary translation product. Cleavage of the C-terminal portion showed to have no significant effect on the basic enzyme properties.

• Biomolecules & Therapeutics
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• 제6권2호
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• pp.204-212
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• 1998

This study was conducted to evaluate the repeated dose toxicity of DA-1131/betamipron, newly developed carbapenem antibiotic, in rats. DA-1131/ betamipron was administered intravenously once a day for 4 weeks to 10 males and 10 females per group at the doses of 0(control),40, 160 and 640 mg/kg. Throughout the study period, all rats survived. The administration of DA-1131/betamipron induced soft stool or diarrhea in rats of both sexes receiving 160 or 640 mg/kg. The water consumption was increased with a statistical significance in 640 mg/kg during observation period. At the end of administration, hematological and serum biochemical examination showed no toxicological changes in DA-1131/betamipron treated groups compared with control group. Histopathologic examination revealed inflammatory cell infiltration, tubular dilatation and focal necrosis of kidney in two males and three females in 640 mg/kg. On the basis of these results, the noobserved-adverse-effect-level of DA-1131/betamipron was estimated to be 40 mgtg under the present test condition.

• 생명과학회지
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• 제7권2호
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• pp.127-133
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• 1997

Phytase(myo-inositol hexakisphosphate phosphohydro-lase;EC 3.1.38)활성은 소화기관인 소장 점막에서나 나타났고, 그 외 다른 장기에서는 alkaline phosphatase활성만을 측정할 수 있었다. 그리고 anti-90kDa phytase항혈청을 이용한 면역조직학적 조사 결과 소장 이외의 장기에서는 본 효소의 단백질 밴드가 검출되지 않았다. 이와 같은 결과 phytase는 소장에만 특이적으로 존재하며, 소장 특이적 효소의 생리적 역할로서 Phytic acid(inositol=hexakisphos-phate)를 가수분해한다. 흰쥐의 성장 발육과 더불어 phytase의 활성은 증가한다. 출생 후부터 이유기 전까지는 70kDa phytase외에도 90kDa phy-tase가 출현한다. 이 90kDa phy-tase는 이유기에 합성되는 것으로 추정된다.

• 한국동물학회지
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• 제35권3호
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• pp.322-331
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• 1992

To investigate the svnthyesis of muscle proteins during differentiation of chicken myoblast, cvtosolic and membrane fractions were used for both sodium dodecvl sulfate polvcrylamide gel eBectrophoresis and two-dimensional gel electrophoresis. An extensive cell fusion was observed in 4 day culture. In the protein pattern of the cvtosolic fraction from SDS-PAGE. several protein bands including 250 kDa and 46 kDa showed remarkable changes during culture. the protein of 46 kDa was the most prominent one ann its optical density was the highest in 5 day culture (OD = 1.30). In the membrane fraction, band of 19.8 kDa showed the highest absorbance with 0.93 OD at 12 hr after initial plating and decreased gradually thereafter to 0.23 in 5 nay culture. From the results of two-dimensional gel electrophoresis of cytosolic fraction, the 46 kDa spot was observed as ko separated forms from culture 2 nary culture, and the sixte of this spot was the largest in 5 nay culture. In the pattern of membrane protein, the extensive appearance of newiv synthesized Proteins was found in a naut culture, but no Prominent spot was observed throughout culture. From the results of the present clay, we found that, during myoblast differentiation, the most prominent proteins were bands of 46 kDa and 19.8 kDa in cvtosolic and membrane fraction, respectively, and the appearance of new proteins was initiated at 48 hr after initial plating, and the 46 kDa protein was predominant in the cytoplasm of late culture in which extensive cell fusion was observed.

• Journal of Photoscience
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• 제9권2호
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• pp.82-85
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• 2002

33kDa extrinsic protein, an important protein in oxygenic photosynthesis, was known to have no fixed configuration in solution. At 20$\^{C}$ and pH 6, 33kDa extrinsic protein showed changes of free energy of -14.6 kJ/mor$\^$-1/ and of standard volume of -120mL/mol, respectively, with increase of hydrostatic pressure, comparatively lower than for most proteins. NBS modification of Trp241 in 33kDa extrinsic protein dramatically changes the secondary protein structure, its affinity to photosystem II as well as photosynthetic oxygen evolution. The relationship between structural change and transport of oxygen, water and proton is deserved a further study.

• Biomolecules & Therapeutics
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• 제3권1호
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• pp.47-53
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• 1995

DA-125, a new anthracycline antitumor antibiotic, was administered at dose levels of 0, 0.2, 0.6 and 1.8 mg/kg/day intravenously to pregnant New Zealand White rabbits from day 6 through 18 of gestation. The does were subjected to the caesarean section on day 28 of gestation. Effects of test agent on general toxicity of does and embryonic development of F1 fetuses were examined. At 1.8 mg/kg, the organ weight for ovary of does was significantly decreased. The decrease in the number of corpus lutea, implantations and litter size, and the increase in the rate of resorptions were also observed. In addition, various types of external, visceral and skeletal malformations occurred in fetuses at an incidence of 7.7, 7.7 and 20.6%, respectively. The results show that the no effect dose levels (NOELs) of DA-125 are 0.6 mg/kg/day for does and F1 fetuses.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.587-601
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• 2013

During the last few decades, it has become evident that the compatibility of the yeast biochemical environment with the ability to process and translate the RNA transcript, along with its capacity to modify a translated protein, are relevant requirements for selecting this host cell for protein expression in several pharmaceutical and clinical applications. In particular, Pichia pastoris is used as an industrial host for recombinant protein and metabolite production, showing a powerful capacity to meet required biomolecular target production levels in high-throughput assays for functional genomics and drug screening. In addition, there is a great advantage to using P. pastoris for protein secretion, even at high molecular weights, since the recovery and purification steps are simplified owing to relatively low levels of endogenous proteins in the extracellular medium. Clearly, no single microexpression system can provide all of the desired properties for human protein production. Moreover, chemical and physical bioprocess parameters, including culture medium formulation, temperature, pH, agitation, aeration rates, induction, and feeding strategies, can highly influence product yield and quality. In order to benefit from the currently available wide range of biosynthesis strategies using P. pastoris, this mini review focuses on the developments and technological fermentation achievements, providing both a comparative and an overall integration analysis. The main aim is to highlight the relevance and versatility of the P. pastoris biosystem to the design of more cost-effective microfactories to meet the increasing demands for recombinant membrane proteins and clinical antibodies for several therapeutic applications.

• 한국임상수의학회지
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• 제31권6호
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• pp.490-494
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• 2014

개의 아토피성 피부염에서 Dermatophagoides farinae의 항원을 immunoblot을 통해 확인하였다. 실험은 D. farinae에 의해 아토피성 피부염이 유발된 16마리와 임상적으로 건강한 10마리 개의 혈청으로 수행하였으며, SDS-PAGE를 실시하여 D. farinae 추출물의 항원들이 15에서 220 kDa 범위 내에 존재하는 것을 관찰하였다. Immunoblot 결과, 임상적으로 건강한 10마리 개의 혈청에서는 단백띠가 보이지 않았으며, D. farinae에 의해 아토피성 피부염이 유발된 16마리 개의 혈청을 이용한 실험에서는 14마리에서 109 kDa, 13마리에서 44 kDa, 11마리에서 177 kDa, 7마리에서 27 kDa, 5마리에서 75 kDa, 3마리에서 15 kDa의 단백띠가 관찰되었다. 즉, D. farinae에 의해 아토피성 피부염이 유발된 개체 중 50% 이상에서 관찰되는 주요 항원은 177 kDa, 109 kDa, 44 kDa 항원이었다. 본 실험과 같이 주요 항원들을 확인하는 이러한 연구는 아토피성 피부염에 대한 새로운 면역 치료의 전략과 제제의 개발에 기여할 수 있을 것으로 생각된다.

• Archives of Pharmacal Research
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• 제22권2호
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• pp.165-172
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• 1999

In the present work , the transport mechanism of a capsaicin derivative, DA-5018, through blood-brain barrier (BBB) has been investigated to evaluate the feasibility of potential drug development. The result of pharmacokinetic parameters obtained from the intravenous injection of plasma volume marker,$[3^H]RSA$ and $[{14}^C]DA-5018$, indicated that both AUC, area under the plasma concentration curve and VD, volume of distribution in brain of $[3^H]RSA$ agreed with those reported ($1620{\pm}10 $percentage injected dose minute per milliliter (%IDmin/ml) and $12.0{\pm}0.1{\mu}l/g$, respectively). Elimination half-life and AUC of $[{14}^C]DA-5018$is corrected by the PHLC analysis, 19.6$\pm$1.2 min and 7.69$\pm$0.85% IDmin/ml, respectively. The metabolic rate of $[{14}^C]DA-5018$was very rapid. The blood-brain barrier permeability surface area (PS) product of $[{14}^C]DA-5018$ was calculated to be 0.24$\pm$0.05 $\mu$l/min/g. The result of internal carotid artery perfusion and capillary depletion suggested that [14C]DA-5018 pass through BBB with the time increasingly. Investigation of transport mechanism of $[{14}^C]DA-5018$ using agonist and antagonist suggested that vanilloid (capsaicin) receptor did not exist in the BBB, and nutrient carrier system in the BBB has no effect on the transport of DA-5018. In conclusion, despite the fact that penetration of DA-5018 through BBB is significant, the intact drug found in the brain tissue is small because of a rapid metabolism. Therefore, for the central analgesic effect of DA-5018, the method to increase the metabolic stability in plasma and the brain permeability should be considered.

• Journal of Animal Science and Technology
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• 제64권4호
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• pp.792-799
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• 2022

Dopamine (DA) is known to be a key modulator of animal behaviors. Thus, the plasma concentration of DA might be used as a biomarker for the behavioral characteristics of horses. The behavioral characteristics of horses vary depending on the breed, age, and sex. Moreover, the DA receptor genotypes are also related to horse behaviors. Thus, the aim of this study was to investigate the DA concentration variations of horse plasma by breed, age, sex, or genotype of its receptor. The horses were divided by breed into Thoroughbred (n = 13), Pony (n = 9), Warmblood (n = 4), and Haflinger (n = 5). The age variable was divided into three different groups: post-pubertal (2-5 years, n = 6), adult (6-13 years, n = 19), and aged horses (15-24 years, n = 6). The sex variable was divided into geldings (n = 8) and mares (n = 23). Approximately 10 mL of blood was collected, and an ELISA kit was used to measure the plasma concentration of DA. Polymerase chain reaction analysis was performed to identify the genetic variation in the DA D4 receptor gene (DRD4). SPSS statistical software was used for statistical analysis. The DA concentrations in geldings were significantly lower than those in mares. There was no significant difference in DA concentrations among breed and age groups. Horses with the GG and GA genotypes had significantly higher plasma concentrations of DA compared to horses with the AA genotype for the G292A gene. Briefly, the plasma concentration of DA varied depending on the sex and genotype of G292A. These factors should be considered when the concentration of DA is used as a biomarker for the behavioral characteristics of horses. In conclusion, the DA concentration or DRD4 genotype of horse plasma has the potential to be used as a biomarker that can predict the behavioral characteristics of horses.